红芸豆miR395家族成员的鉴定及其与结瘤相关的表达分析  

Identification of miR395 Family Members and Their Nodulation Relevant Expression Analysis in Red Kidney Bean

作　　者：王东梅 孙丽丽 郑江涛 王利祥 郭数进[1] WANG Dongmei;SUN Lili;ZHENG Jiangtao;WANG Lixiang;GUO Shujin(College of Agriculture,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学农学院,山西太谷030801

出　　处：《山西农业科学》2023年第10期1170-1177,共8页Journal of Shanxi Agricultural Sciences

基　　金：国家自然科学基金项目(32241046);山西省科技重大专项(202101140601027);山西省科技创新人才团队项目(202204051001020);山西省基础研究计划项目(20210302123365)。

摘　　要：microRNA395是高等植物保守的miRNA家族,主要通过靶向低亲和力硫酸盐转运蛋白(AST)和ATP磺酰化酶(APS)来响应植物的硫饥饿反应,硫饥饿会显著影响固氮植物的结瘤共生固氮过程。为解析miR395在豆科植物结瘤共生固氮中的作用,对红芸豆miR395家族成员进行生物信息学预测,并对pvu-miR395在红芸豆不同组织和根瘤菌侵染不同时间段进行差异表达分析。结果显示,在全基因组水平上鉴定出8个pvu-miR395的家族成员,位于第2、3号染色体上。pvu-miR395家族成员成熟序列和星标序列高度保守,它们的前体序列均可生成稳定的二级结构。进一步对芸豆与大豆、苜蓿、拟南芥、水稻miR395家族成员进行系统进化分析,可将miR395家族成员分为GroupⅠ~Ⅳ4个亚家族。顺式作用元件分析发现,该家族成员可能参与植物激素、胁迫、生长和发育等过程;qRT-PCR差异表达分析发现,pvu-miR395家族成员均在根瘤中表达量最高,具有组织特异性,且pvu-miR395的表达量随根瘤菌处理时间延长呈现先升高后降低的趋势,显著抑制根瘤菌的侵染。靶基因预测表明,硫酸盐转运蛋白和硫酸腺苷酰基转移酶可能是pvu-miR395的靶基因。MicroRNA395 is a conserved miRNA family in higher plants,which mainly responses to sulfur starvation by targeting low affinity sulfate transporter(AST)and ATP sulfonylases(APS).Sulfur starvation significantly affects the process of nodulation and symbiotic nitrogen fixation of nitrogen fixing plants.To elucidate the role of miR395 in nodulation and symbiotic nitrogen fixation in leguminous plants,in this study,bioinformatics prediction was performed on the member of pvu-miR395 family members in red kidney bean,and differential expression analysis of pvu-miR395 in different tissues and at different periods of rhizobium infection was conducted.The results showed that eight members of miR395 family were identified in whole genome level,located on chromosomes 2 and 3.The mature and star sequences of miR395 family members in red kidney bean were highly conserved,and their precursor could generate stable secondary structures.Further phylogenetic analysis of miR395 family members in Phaseolus vulgaris,Glycine max,Medicago truncatula,Arabidopsis thaliana,and Oryza sativa were conducted to divide the members into four subfamilies(GroupⅠ-Ⅳ).cis-acting element analysis revealed that the family members might be involved in plant hormones response,stress response,plant growth,and development.qRT-PCR differential expression analysis showed that pvu-miR395 family members had the highest expression levels in nodules with tissue specificity.The expression of pvu-miR395 increased first and then decreased with the treatment time of rhizobium,significantly inhibiting rhizobium infection.Target gene prediction showed that sulfate transporter and adenylyl sulfate transferase might be the target genes of pvu-miR395.

关 键 词：红芸豆 miR395 生物信息学分析 靶基因 结瘤 表达分析 

分 类 号：S643[农业科学—蔬菜学]